Fig. 4: RNA degrading activity and anti-viral activity of C30-based RIBOTACs. | Nature Communications

Fig. 4: RNA degrading activity and anti-viral activity of C30-based RIBOTACs.

From: Chemical-guided SHAPE sequencing (cgSHAPE-seq) informs the binding site of RNA-degrading chimeras targeting SARS-CoV-2 5’ untranslated region

Fig. 4

a Synthesis of C30-based RIBOTACs using conjugation sites on rings A or E of C30. b Comparison of two RLR moieties in the RIBOTAC modality using the in vitro RNase L degradation assay with purified SL5 RNA. RNase L and compounds were preincubated at 4 °C for 12 h to induce dimerization of RNase L. Cyanine 5 (Cy5)-labeled SL5 RNA was then added, and the mixture was incubated at 22 °C for an additional 2 h. RNA degradation products were analyzed by polyacrylamide gel electrophoresis (PAGE) using the red fluorescence channel (right), while the RNA ladder was visualized with SYBR Safe stain and imaged using the green fluorescence channel (left). Experiments were performed three times independently, yielding similar results. c Cellular activity of RIBOTACs in SARS-CoV-2 5’ UTR expressing cells. d Inhibitory effect of RIBOTAC C64 in SARS-CoV-2 infected A549 cells. The cytotoxicity of the compound was also evaluated. The dose-response curves are representative of three independent measurements (n = 3). Data are presented as mean values ± SD. Source data are provided in the Source Data file.

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